Genetic resources of Cannabis sativa L. at the Institute of Natural Fibres and Medicinal Plants in Poznań

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1 Genetic resources of Cannabis sativa L. at the Institute of Natural Fibres and Medicinal Plants in Poznań Grażyna Mańkowska*, Lidia Grabowska Institute of Natural Fibres and Medicinal Plants Wojska Polskiego 71B Poznań, Poland *corresponding author: S u m m a r y Hemp (Cannabis sativa L.) species belongs to the Hemp family (Cannabaceae), Cannabis genus. Hemp is considered as a species easily adapting to the changing environment conditions. It is characterized by plentiful of forms showing considerable biological and morphological differences and different economical value. Development of such big differentiation took place under the effect of geographical, climatic and soil conditions as well as human activity which aimed at developing and stabilizing important qualities of hemp. The collection of hemp genus held at the Institute of Natural Fibres in Poznań contains about 150 accessions. The collection holds both mono- and dioecious forms of hemp from zones represented by different soil and climatic conditions, local ecotypes and breeding lines with stabilized genotype that are exceptional in respect of one parameter, for instance Δ9THC, yield, quality of fiber, composition of fatty acids, etc. In 2000, the INF genetic resources were transferred to new facilities providing excellent conditions of storage. The long-term storage of seeds requires low air temperature, low seed moisture content (7 8%) and proper relative air humidity (below 55%). The air-dried seeds (7% of moisture content) are packed hermetically in a thermo-sealed aluminum foil and sent to core collection stored in a chamber at +4.0 o C or to reserve bank where the 5-g samples are stored for a long time. Key words: genetic resources, monoecious hemp, dioecious hemp, accession, collection, accession evaluation

2 INTRODUCTION Genetic resources of Cannabis sativa L. at the Institute of Natural Fibres and Medicinal Plants in Poznań Hemp (Cannabis sativa L.) belongs to the Cannabaceae family, Cannabis genus. Hemp plants are considered to be readily adapting to changing environmental conditions. Hemp is characterized by a multitude of diverse forms of biological and morphological traits and the different economic value [1, 2]. The formation of such a large difference occurred as a result of geographical, climatic and soil conditions and human activities, which aimed at establishing and maintaining important features of commercial hemp. Currently, the Institute of Natural Fibres and Medicinal Plants holds a collection consisting of 150 genotypes, including monoecious and dioecious forms of hemp, from different regions of the world, local ecotypes and breeding lines with a stable genotype, unique in terms of at least one parameter, such as the THC content, yield, fiber quality, fatty acids composition etc. Evaluation work is carried out on collected objects, aimed at renewal and multiplication of seed and allows possession to carry out the work of valorization. For test objects, in addition to distinctive features, morphological and biological traits and use value were determined. This data is valuable for breeders in selecting parental components. 179 MATERIAL AND METODS Renewal of sowing and the multiplication of genotypes. Propagation and renewal of sowing material in the collection is conducted at the Experimental Station of Institute of Natural Fibres and Medicinal Plants in Pętkowo (Wielkopolska province). Due to dioeciousness, allogamy and anemophily of hemp, the work related to the propagation and renewal of sowing material requires spatial isolation. Small batches of seed are planted in isolated pots located in the vegetation hall and larger batches of seed in isolated chambers, greenhouses and with the isolation in the field. In case of reproduction of monoecious forms, it is necessary to remove the male specimen during flowering. Another problem in case of cannabis is that the seed very fast loses its germination capacity [3]. Therefore, the seeds of germination capacity higher than 80% are controlled every 3 years, those with germination capacity lower than 80% every year. Determination of the economic value A series of comparative field experiments was conducted in order to determine the economic value of the stored genotypes in accordance with the agronomical recommendations of the hemp fiber. Annually, a bulk of objects are tested in two repetitions of 300 seeds on 1.5 m 2 plots. Vol. 55 No

3 G. Mańkowska, L. Grabowska 180 During vegetation several observations are carried out e.g. development phases of hemp, lodging and diseases as well as pests and plant number per unit area. During the flowering inflorescences are sampled in order to determine a hallucinogenic substances. The following tests were performed in the laboratory: measurements of morphological traits: total length, stem length and diameter; 1000 seed mass, determined according to ISTA [4]; determination of fiber content in straw, according to the method of osmotic degumming developed at INF [5]; cellulose content, according to PN -92/pn-50092; content of hallucinogenic substances using gas chromatograph in accordance with EU Council Regulation (EC) No 1251/1999, Annex 5, Art. 7b, annex 1. Białobrzeskie variety serves as a reference cultivar the first Polish monoecious fibrous hemp variety, registered in 1968 in the National Register of COBORU, which contains less than 0.2% of hallucinogenic substances, is a Central European form adapted to Polish climate and soil conditions. The vegetation period of this variety is days, plant height (depending on cultivation technology) cm, seed yield (depending on the purpose of growing t/ha), straw yield about 10 t/ha fiber yield t/ha [6]. RESULTS AND DISCUSSION Cellulose fiber content Processing technology of raw material was carried out at the Laboratory Evaluation of Textile Raw Materials, Institute of Natural Fibers and Medicinal Plants. The results of the determination of fiber and cellulose content of hemp straw is shown in table 1. Fiber content in the sites surveyed in was highly variable and ranged from 12% for the Chinese variety He Bei to 29.2% for the Hungarian variety Uniko B. The reference variety contains 26.6% of fiber. This value is exceeded only by 4 varieties: Uniko B, Jus 8, Juso 15, Chamaeleon and Juso 11. The lowest fiber content was found in a group of accessions with the longest growing season He Bei, and varieties with the shortest period of vegetation Finola. These results confirm the literature data that indicate that at the flowering phase of growth of biomass is small but biological processes in stems lead to the rise in fiber content [7, 8]. The cellulose content in hemp straw demonstrated less variability than the fiber content. The lowest cellulose content was found in the Bulgarian variety Silistrenskie (37.7%) and the highest in the Ukrainian variety Jus 8 (46.2%), while this figure was just 0.3% higher than for the reference.

4 Genetic resources of Cannabis sativa L. at the Institute of Natural Fibres and Medicinal Plants in Poznań Ta b l e 1. The content of fiber in selected varieties of hemp collected in the gene bank in cultivar average fibre content [%] cellulose content [%] Zenit Juso Futura Chamaeleon Diana Uniko B Silistrenskie Fedora Finola Juso K Dnieprowska Fibrimon Zolotonoska Gigantheus He Bei Jus Białobrzeskie seed weight Special studies were conducted on the quality of selected hemp species. The indicator of seed size is the measure of thousand seed mass. With regard to the useful characteristics as well as morphological and biological differences, hemp can be divided into three groups, namely Northern, Southern and Central European ones. Thousand seed mass for each hemp group is different. Average weight of 1000 seeds for Southern hemp is g, intermediate hemp g and 8 12 g for wild hemp [9]. Varieties such as: Silistrenskie, Chamaeleon, Zenit exceeded significantly the MTS value of the Białobrzeskie variety. Finola variety of all tested cultivars is characterized with the lowest value of the 1000 seed weight (tab. 2). THC content The contents of narcotic substances in cannabis Δ9 THC (tetrahydrocannabinol) is the result of interaction of both genetics and environmental conditions. Genetic predisposition, environmental conditions, different stages of development and plant fragments can cause significant fluctuations in THC content (from to 0.1%, see tab.2). Vol. 55 No

5 G. Mańkowska, L. Grabowska 182 THC content has been the highest for Finola variety, which value in our climatic conditions reaches 0.2% and therefore can be considered as a drug type variety. Ukrainian varieties reached the lowest value of THC [10]. Ta b l e 2. Mass of a thousand seeds THC content (%) in hemp cultivars collected in the Institute of Natural Fibres and Medicinal Plants in cultivar origin mass of 1000 seeds [g] THC [%] Zenit Romania Juso 11 Ukraine Futura 77 France Chamaeleon The Netherlands 18.0 Diana Russia Uniko B Hungary Silistrenskie Russia Fedora 17 France Finola Finland Juso 15 Ukraine K Poland Dnieprowska Ukraine Fibrimon 24 France Zolotonoska 26 Ukraine Gigantheus Italy He Bei China Jus 8 Ukraine Białobrzeskie Poland CONCLUSIONS 1. Finola (30.05) and He Bei (1.09) begin flowering at the earliest among the dioecious varieties of hemp. When it comes to monoecious varieties, the earliest blooming were Juso11, Juso15 (8.07), and the latest was Futura 77 (Tab. 3). 2. The highest fiber content (29.16%) was reported for the Hungarian variety Uniko B, while the lowest (12.00%) for the Chinese variety He Bei. 3. The highest content of cellulose has been reported for Jus 8 (46.2%), and the lowest for Silistrenskie (37.74%). 4. The examined accessions were characterized by the mass of 1000 seed from 19.0 g (Jus 8) to 10.3 g (Finola). 5. Finola variety was characterized by higher content of THC in comparison with the standard variety.

6 Genetic resources of Cannabis sativa L. at the Institute of Natural Fibres and Medicinal Plants in Poznań Ta b l e 3. Morphological parameters of some hemp cultivars from Institute of Natural Fibres and Medicinal Plants collection cultivar form beginning of the flowering full maturity Zenit monoecious Juso 11 monoecious Futura 77 monoecious Chamaeleon dioecious Diana dioecious Uniko B dioecious Silistrenskie dioecious Fedora 17 monoecious Finola dioecious Juso 15 monoecious K-195 dioecious Dnieprowska monoecious Fibrimon 24 monoecious Zolotonosha26 monoecious He Bei dioecious Can.gigantheus dioecious Białobrzeskie monoecious REFERENCES 1. Białousowa J, Bartosik A, Kurhański M, Nagórski A, Tumalewicz B. Konopie, Rośliny włókniste [Fibrous Plants]. Warszawa 1958: Bosca I, Karus M. Der Hanfanbau. Heidelberg 1997: Ziemnicki Z, Wierzchowiecka K. Wpływ przechowywania nasion konopi w różnych opakowaniach na zachowanie zdolności kiełkowania. Poznań 1984; 29: Duczmal K, Tucholska H. Nasiennictwo. Tom 1. Warszawa 2000: Allam A, Kozłowski K, Konczewicz W. Degumming of fibrous plants based on osmotic pressure phenomenon. Proceedings of International Conference of FAO European Research Network on Flax and Allied Fibre Plants for Humanity Welfare, Cairo Grabowska L, Jaranowska B, Baraniecki P, Tymków J. The Results of Hemp Breeding in Poland. Poznań 1998; 2: Jaranowska B. Wpływ stopnia dojrzałości konopi jednopiennych uprawianych na glebach mineralnych na wydajność i jakość włókna. Prace IPWŁ. Poznań 1968; 15: Chocianowicz W, Cierpucha W. Kurhański M, Szałkowski Z, Waśko J.Poradnik brakarza włókna lnianego i konopnego. Poznań Bytnerowicz H. Konopie. Warszawa 1968: Grabowska L, Mańkowska G, Baraniecki P. Zasoby genowe Cannabis sativa L. w Instytucie Włókien Naturalnych w Poznaniu. Zeszyty Problemowe PNR Warszawa 2004; 497:53-7. Vol. 55 No

7 G. Mańkowska, L. Grabowska 184 Zasoby genowe konopi Cannabis sativa L. w Instytucie Włókien Naturalnych i Roślin Zielarskich w Poznaniu Grażyna Mańkowska*, Lidia Grabowska Instytut Włókien Naturalnych i Roślin Zielarskich ul. Wojska Polskiego 71B Poznań *autor, do którego należy kierować korespondencję: biotech@inf.poznan.pl S t r e s z c z e n i e Konopie (Cannabis sativa L.) są gatunkiem należącym do rodziny konopiowatych Cannabaceae, rodzaju Cannabis. Uważane są za roślinę łatwo adaptującą się do zmiennych warunków środowiskowych. Charakteryzuje się mnóstwem form o zróżnicowanych cechach biologicznych i morfologicznych oraz o różnej wartości ekonomicznej. Wytworzenie się tak dużych różnic wystąpiło jako efekt działania warunków geograficznych, klimatycznych i glebowych oraz działalności człowieka, która miała na celu stworzenie i utrzymanie ważnych cech konopi. Kolekcja rodzaju konopi w Instytucie Włókien Naturalnych i Roślin Zielarskich w Poznaniu zawiera około 150 obiektów. Zawiera zarówno jedno- i dwupienne formy charakterystyczne dla różnych warunków glebowo-klimatycznych, lokalne ekotypy i linie hodowlane ze stabilnym genotypem, wyjątkowym pod względem jednego parametru, np. zawartości THC, plonowania, jakości włókna, składu kwasów tłuszczowych itp. W 2000 r. zasoby genowe Instytutu Włókien Naturalnych zostały przeniesione do nowego miejsca, które zapewnia doskonałe warunki przechowywania nasion. Długoterminowe przechowywanie wymaga niskiej temperatury, niskiej wilgotności nasion (7 8%) oraz odpowiedniej względnej wilgotności powietrza (poniżej 55%). Osuszane powietrzem nasiona (7% wilgotności) są zapakowane hermetycznie w zgrzewaną folię aluminiową i umieszczane w kolekcji bazowej (przechowywanie w komorze w temperaturze +4,0ºC) lub banku rezerwowym próbki 5-gramowe są przechowywane przez dłuższy czas. Słowa kluczowe: zasoby genowe, konopie jednopienne, dwupienne, obiekty, kolekcja, ocena obiektów